Brief Summary
This video explains enzyme immobilization, focusing on a practical example using yeast cells (containing the enzyme sucrase) trapped in a sodium alginate gel. It covers the immobilization process, testing the application of the immobilized enzyme, comparing it to a control (free yeast), and discussing the advantages of enzyme immobilization. The video also provides real-world examples of enzymes in use, such as pectinase and glucose isomerase.
- Enzyme immobilization involves fixing enzymes to each other or an inert material.
- Trapping enzymes in a gel (like sodium alginate) is a common immobilization method.
- Immobilized enzymes offer advantages like stability, reusability, and purer products.
The video introduces enzymes as biological catalysts, specifically focusing on sucrase found in yeast cells. Sucrase acts on sucrose (the substrate) to produce glucose (the product). The practical demonstration involves immobilizing yeast cells to study the application of the enzyme.
Enzyme immobilization is defined as fixing enzymes to each other or to an inert material. In this experiment, yeast cells are immobilized by trapping them in a sodium alginate gel. The gel is prepared by mixing sodium alginate and distilled water, while the yeast is mixed separately with distilled water. The yeast solution is then added to the sodium alginate gel and mixed thoroughly. This mixture is drawn into a syringe and slowly dropped into a calcium chloride solution, forming beads. The calcium chloride hardens the beads, trapping the yeast cells within the alginate gel. The beads are left to harden for 10 minutes and then washed with distilled water to remove any free yeast cells from the surface.
Besides trapping in a gel, other immobilization methods exist. Physical methods include attaching enzymes to ceramic beads. Chemical methods involve bonding enzymes to physical supports or to each other.
To test the application of the immobilized enzyme, the beads are transferred to a separating funnel with a straw to prevent clogging. Sucrose solution (the substrate) is added, and the solution is tested for glucose using glucose test strips every 2 minutes until a positive result is obtained.
A control experiment is conducted using free yeast in a separating funnel with sucrose solution. The solution is tested immediately and every 2 minutes for glucose until a positive result is obtained, allowing for a comparison with the immobilized yeast.
The results show that it takes longer for the immobilized yeast to produce a positive glucose result compared to the free yeast. This is because the sucrose has to diffuse through the pores of the gel to reach the yeast cells, and the product has to diffuse out. However, the product from the immobilized yeast is much clearer than the product from the free yeast, which is mixed with yeast cells.
The advantages of enzyme immobilization include stabilizing the enzyme, allowing for reuse of the enzymes, and obtaining a purer product that does not require extensive purification.
Pectinase is used in food processing to extract and clarify fruit juices. Glucose isomerase is used in industry to convert glucose to fructose, reducing the need to add more sugar to products.
The practical demonstration of enzyme immobilization ties into broader topics such as bioprocessing, single-cell proteins, and bioreactors.
Summarize any YouTube video in seconds
Get summaries like this one for any video, in your language, with the free Summ app.
Transcript
Full transcript of “Enzyme Immobilisation-Leaving Cert Biology”. Click a timestamp to open the video at that moment. Need another video? Use the YouTube transcript generator.
0:00 mzy immobilization and testing its application this practicle is all about mzes it's all about those biological catalysts that are protein in nature and folded into that three-dimensional globular shape in this practical yeast was used and yeast cells contain the mzy sucra sucrose solution was also used the sucrose acted as the substrate and the substrate was changed into glucose which was our product so what exactly does immobilization mean the correct definition is that immobilized enzymes are enzymes that are fixed to each other or some other inert material we immobilized our yeast cells by trapping them in a gel and that gel
0:45 was sodium alginate the gel was prepared by mixing sodium alginate and distilled water in a beaker the yeast was prepared by mixing it with distilled water in a separate Beaker the yeast solution was added to the sodium alginate so it was added to the gel and this mixture was mixed very well separately a calcium chloride solution was prepared The Next Step involved drawing up the yeast and sodium alginate mixture into a syringe very slowly small amounts of the yeast and sodium alginate mixture were dropped from the syringe into the beaker of calcium chloride solution as this gel and yeast mixture hit the calcium chloride solution perfect beads were formed and the function of the calcium chloride solution is to harden the beads
1:33 when all the yeast and sodium alginate was dropped from the syringe into the calcium chloride solution we let the beads sit for 10 minutes to harden the beads were removed from the calcium chloride solution and washed with distilled water washing with distilled water removes any possible free yeast that might be on the outside of the beads so that was how we immobilized our yeast cells in a gel here's a rough diagram of one of those beads so you have the alic gel you have the pores and you have the yeast cells inside do we have to use a gel no there are other methods of immobilization the first way of immobilizing would be physical methods so just as we did trapping in a gel or another way would be to attach the enzymes to ceramic beads chemical
2:20 methods of immobilization would be to bond the enzymes to physical supports or to bond them to each other now that we have immobilized our enzyme we have to test its application what's it like to use the first thing we did was we transferred our beads to a separating funnel and we had a straw in the funnel to prevent the tap from getting clogged then we added our substrate which is the sucrose solution we immediately tested with a glucose test strip and then every 2 minutes afterwards the solution was tested the process was continued until the glucose test strip changed color to indicate the presence of glucose so all good scientific experiments need something for comparison so they need a control our control was free yeast in
3:07 the separating funnel and then we added our sucro solution we tested straight away using a glucose test strip and then every 2 minutes until a positive result for glucose was obtained the results let's discuss the immobilized yeast first it took much longer for a positive result for glucose to appear but the product was much clearer why did it take so long for a positive result well the sucrose has to make its way in through the pores of the gel and then has to mix with the yeast cells and the product has to find its we out of the pores that takes time in contrast the free yeast tested positive for glucose much faster but the product was mixed with yeast what is the benefit or what are the advantages of immobilization the first is that
3:53 immobilization stabilizes the enzyme the second is that the enzymes can be reused the third is that you get a purer product a product that does not have to go through purification be sure you can give some examples of mzes in use and consult your textbooks for Relevant examples but here are a few pectinase is an mme that's used in food processing it's used to extract juices particularly from apples and oranges and it's also used to clarify to remove the cloudiness from fruit juices glucose isas is used in industry to convert glucose to the sweeter tasting fructose this avoids you having to add more sugar to your product so that was the Practical on MZ immobilization it ties in nicely with revising your enzymes checking out
4:39 bioprocessing single cell proteins and bioreactors good luck [Music]
